Two functionally distinct myeloid dendritic cell subpopulations are present in bovine blood.

Immature myeloid (m)DCs circulating in the blood of cattle have been defined as lineage negative (Lin(-))MHCII(+)CD11c(+)CD205(+) cells. Lin(-)MHCII(+)CD11c(+)CD205(+) mDCs (0.2% blood mononuclear cells) isolated from bovine blood were heterogeneous in cell size and CD205 expression. Using highspeed cell sorting, Lin(-)MHCII(+)CD11c(+)CD205(+) DCs were sorted into CD205(Hi) and CD205(Lo) subpopulations which were phenotypically distinct and differed significantly (P<0.01) in TLR gene expression. CD205(Hi) and CD205(Lo) mDCs were more efficient in macropinocytosis than monocytes and expressed no or little detectable non-specific esterase activity. CD205(Lo) mDCs efficiently activated purified allogeneic T cells and the addition of TLR agonists did not significantly alter this antigen presentation capacity. T cell activation by CD205(Lo) mDCs was associated with differential up-regulation of CD40, CD80, CD86 and TGFß1 gene expression when compared to CD205(Hi) mDCs. In conclusion, two phenotypically and functionally distinct CD11c(+)CD205(+) mDCs were isolated from blood that had an equal capacity to acquire antigen but markedly different capacities to activate T cells.

Age-related changes in the distribution and frequency of myeloid and T cell populations in the small intestine of calves.

Mucosal dendritic cells (DCs) play a key role in discriminating between dietary antigens, commensal microflora and pathogens but little is known regarding age-related changes in mucosal DC populations. We analyzed lymphoid and myeloid populations within the epithelium and lamina propria (LP) of the ileum and jejunum of weaned calves (6 months old) and compared their frequency and distribution with newborn calves (3-5 weeks old). CD4, CD8 and γδ TcR T cells and CD11c(Hi)MHC Class II(+) myeloid cell frequency were significantly different when comparing ileum and jejunum of weaned calves. In particular, the number of CD8 and γδ TcR T cells, and CD11c(Hi)CD14(+) macrophages was significantly greater in the ileum but CD11c(+) and CD11b(+) myeloid cell distribution was similar throughout the mucosal epithelium of the small intestine. Furthermore, significant age-related changes were apparent when comparing the frequency and abundance of mucosal leukocyte subpopulations in newborn and weaned calves. Total mucosal leukocytes (CD45(+)) increased significantly with age in both ileum and jejunum and much of this increase was attributed to mucosal T cells (CD3(+)). In particular, CD4 T cells and NK cells increased significantly in the jejunum and CD8, and γδ TcR T cells increased significantly with age throughout the small intestine. In contrast, CD11c(Hi)MHC Class II(+) myeloid cells remained numerically unchanged with age but DCs (CD13(+), CD26(+), CD205(+)) were enriched and macrophages (CD14(+), CD172a(+)) were depleted in older animals. Therefore, regional differences between ileal and jejunal mucosal leukocytes changed with age and there was also a marked age-dependent change in the composition of mucosal myeloid cells. These observations have significant implications for host responses to both pathogens and commensal microflora.

Mucosal dendritic cell subpopulations in the small intestine of newborn calves.

Mucosal dendritic cell development in the newborn is poorly understood despite evidence that distinct DC subpopulations populate individual mucosal surfaces. Therefore, we investigated DC phenotype and distribution in the small intestine of newborn calves. DC phenotype was analyzed using flow cytometry and DC distribution was investigated with immunohistochemistry. Purification of CD11c(Hi)MHC Class II(+) cells confirmed CD11c defined myeloid cells and a comparison of neonatal blood and intestine revealed distinct mucosal DC subpopulations. CD11c(Hi)CD14(+) cells were significantly more abundant in newborn ileum versus jejunum and CD335(+) NK cells were the only lymphoid population significantly different in ileum versus jejunum. Immunohistochemistry revealed unique patterns of myeloid cell distribution within the mucosal epithelium, lamina propria, and submucosa. CD11c(+) cells were present within the jejunal but absent from the ileal intraepithelial compartment. In contrast, CD11b(+) cells were present within the ileal but absent from the jejunal intraepithelial compartment. In conclusion, the neonatal small intestine is populated by diverse myeloid subpopulations and significant differences in regional distribution are established early in life. These observations may have significant implications for the response of the newborn to both commensal microflora and enteric pathogens.